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Alexa Fluor™ 647 Mouse Anti-Human CD14
Alexa Fluor™ 647 Mouse Anti-Human CD14
Multiparameter flow cytometric analysis of CD14 expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD14 antibody (Cat. No. 568242/568243; Right Plot). After staining, the erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of CD14 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Multiparameter flow cytometric analysis of CD14 expression on Human peripheral blood leucocyte populations. Human whole blood was stained with either Alexa Fluor™ 647 Mouse IgG1, κ Isotype Control (Cat. No. 565571; Left Plot) or Alexa Fluor™ 647 Mouse Anti-Human CD14 antibody (Cat. No. 568242/568243; Right Plot). After staining, the erythrocytes were lysed with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899). The bivariate pseudocolor density plot showing the correlated expression of CD14 (or Ig Isotype control staining) versus side-light scatter (SSC-A) signals was derived from gated events with the forward and side-light scatter characteristics of viable leucocyte populations. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ software.
Product Details
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BD Pharmingen™
monocyte differentiation antigen CD14; LPS receptor; LPS-R
Human (QC Testing)
Mouse IgG1, κ
Human Peripheral Blood Monocytes
Flow cytometry (Routinely Tested)
5 µl
929
Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
RUO


Preparation And Storage

Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed.

Recommended Assay Procedures

BD® CompBeads can be used as surrogates to assess fluorescence spillover (Compensation). When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.  However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls. It is strongly recommended that when using a reagent for the first time, users compare the spillover on cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
  5. An isotype control should be used at the same concentration as the antibody of interest.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  8. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
  11. For U.S. patents that may apply, see bd.com/patents.
568242 Rev. 2
Antibody Details
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63D3.rMAb

The 63D3.rMAb is a recombinant monoclonal antibody that specifically recognizes human CD14 which is also known as Monocyte differentiation antigen CD14, Myeloid cell-specific leucine-rich glycoprotein, or LPS Receptor (LPS-R). CD14 is expressed as an ~55 kDa glycophosphatidylinositol (GPI)-linked cell surface glycoprotein. CD14 is highly expressed on most mature monocytes and macrophages, and at lower levels on neutrophils. CD14 is also expressed on some dendritic cells (DC) and Langerhans cells. CD14 binds to bacterial cell wall components including lipopolysaccharide (LPS) bound to lipopolysaccharide-binding protein (LBP). CD14 serves as a co-receptor for certain signaling Toll-like receptors (TLRs) for the cellular detection and response to LPS as well as other pathogen-associated molecular patterns (PAMPs). Ligand-bound CD14 plays a role in stimulating various myeloid cells to produce pro-inflammatory cytokines and mediators and to upregulate the expression of cell surface adhesion molecules. Soluble forms of CD14 (sCD14) are reportedly released by cells into the blood or other bodily fluids. Soluble CD14 can regulate cellular responses to LPS and confer LPS-responsiveness to cells that do not ordinarily express CD14.

568242 Rev. 2
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor™ 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 670-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
568242 Rev.2
Citations & References
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View product citations for antibody "568242" on CiteAb

Development References (4)

  1. Devitt A, Moffatt OD, Raykundalia C, Capra JD, Simmons DL, Gregory CD. Human CD14 mediates recognition and phagocytosis of apoptotic cells.. Nature. 1998; 392(6675):505-9. (Clone-specific: Blocking, ELISA, Functional assay, Western blot). View Reference
  2. Rosenberg SA, Ligler FS, Ugolini V, Lipsky PE. A monoclonal antibody that identifies human peripheral blood monocytes recognizes the accessory- cells required for mitogen-induced T lymphocyte proliferation.. J Immunol. 1981; 126(4):1473-7. (Immunogen: Bioassay, Flow cytometry, Fluorescence activated cell sorting). View Reference
  3. Ugolini V, Nunez G, Smith RG, Stastny P, Capra JD. Initial characterization of monoclonal antibodies against human monocytes.. Proc Natl Acad Sci USA. 1980; 77(11):6764-8. (Immunogen: Cytotoxicity, Flow cytometry, Radioimmunoassay). View Reference
  4. Wu Z, Zhang Z, Lei Z, Lei P. CD14: Biology and role in the pathogenesis of disease.. Cytokine Growth Factor Rev. 2019; 48:24-31. (Biology). View Reference
View All (4) View Less
568242 Rev. 2

 

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.